Dog skin parasite load, TLR-2, IL-10 and TNF-α expression and infectiousness.

Visceral leishmaniosis is a zoonotic disease that is transmitted by Lutzomyia longipalpis sandflies. Dogs are the main peri-urban reservoir of the disease, and progression of canine leishmaniosis is dependent on the type of immune response elaborated against the parasite. Type 1 immunity is characterized by effective cellular response, with production of pro-inflammatory cytokines such as tumour necrosis factor alpha (TNF-α). In contrast, Type 2 immunity is predominantly humoral, associated with progression of the disease and mediated by anti-inflammatory cytokines such as interleukin 10 (IL-10). Although seemly important in the dynamics of leishmaniosis, other gene products such as toll-like receptor 2 (TRL-2) and inducible nitric oxide synthase (iNOS) exert unclear roles in the determination of the type of immune response. Given that the dog skin serves as a micro-environment for the multiplication of Leishmania spp., we investigated the parasite load and the expression of TLR-2, iNOS, IL-10 and TNF-α in the skin of 29 infected and 8 control dogs. We found that increased parasite load leads to upregulation of TLR-2, IL-10 and TNF-α, indicating that abundance of these transcripts is associated with infection. We also performed a xenodiagnosis to demonstrate that increased parasitism is a risk factor for infectiousness to sandflies.

Blood-brain barrier disruption during spontaneous canine visceral leishmaniasis.

Visceral leishmaniasis is a complex disease caused by Leishmania infantum, and in dogs, besides the classical symptoms, there are descriptions of inflammatory alterations in the brain. Brain inflammation is a strictly controlled process, and as the brain counts on the efficiency of the blood-brain barrier (BBB), we aimed to assess BBB integrity in dogs with spontaneous visceral leishmaniasis. Therefore, we evaluated markers in the cerebrospinal fluid (CSF) and in brain tissue related to BBB disruption and brain inflammation. Elevated albumin quota revealed BBB breakdown, corroborated by increased concentrations of anti-Leishmania antibodies in the CSF. In the brain, albumin and IgG staining formed halos around blood vessels, a classical indicator of BBB leakage. Soluble IgG was also detected in the choroid plexus and ependyma, and in these structures, IgG stained random resident cells. IgG(+) cells and Fcγ-RI(+) cells were identified in the choroid plexus, ependyma and perivascular in the brain parenchyma. The data support the occurrence of BBB disruption in dogs with spontaneous visceral leishmaniasis, and IgG as a key molecule that is capable of initiating and/or maintaining the inflammatory stimuli in the nervous milieu and the CSF as an important disseminator of inflammatory stimuli within the CNS.

Compartmentalized gene expression of toll-like receptors 2, 4 and 9 in the brain and peripheral lymphoid organs during canine visceral leishmaniasis.

Visceral leishmaniasis is an important parasitic disease that affects humans and animals. The response against the protozoan involves the interaction of both innate and adaptive branches of the immune system, and an important immune sensor is represented by the toll-like receptor (TLR) family. Here, we investigated the pattern of TLR-2, TLR-4 and TLR-9 gene expression in different compartments (brain, choroid plexus, spleen and lymph node) of dogs naturally infected with Leishmania infantum. Gene expression of the TLRs varied according to the compartment evaluated. In the brain, there was only an upregulation of TLR-2, whereas in the choroid plexus, TLR-2 and TLR-9 were both upregulated. Further, the peripheral lymphoid organs (spleen and lymph nodes) showed increased TLR-2 and TLR-4 expression. This study provides the first insight about TLR expression in the central nervous system of infected dogs, and gives additional evidence of the compartmentalization of the immune response during visceral leishmaniasis.

Cardiac lesions in 30 dogs naturally infected with Leishmania infantum chagasi.

The hearts of 30 dogs naturally infected with Leishmania infantum chagasi were evaluated histologically and immunohistochemically. Myocardial lesions were detected in all dogs, including lymphoplasmacytic myocarditis (27/30), myonecrosis (24/30), increased interstitial collagen (22/30), lepromatous-type granulomatous myocarditis (7/30), fibrinoid vascular change (3/30), and vasculitis (1/30). The parasite was detected in the hearts of 20 of 30 dogs. The number of parasitized cells correlated with the intensity of the inflammation and with the number of granulomas. The results indicate that cardiac lesions are prevalent in dogs with naturally occurring leishmaniasis even in the absence of clinical signs of cardiac disease.

Levels of matrix metalloproteinase-2 and metalloproteinase-9 in the cerebrospinal fluid of dogs with visceral leishmaniasis.

Inflammation causes increases in the level of matrix metalloproteinases (MMPs), which, in central nervous system (CNS), are associated with neuroinflammation and disruption of blood-brain barrier. Analysis of cerebrospinal fluid (CSF) is pivotal for detecting diseases in CNS and, although a specific diagnosis may not be achieved, this analysis is helpful to confirm the diagnosis or to rule out relevant differential diagnoses. This study examined the levels of MMP-2 and MMP-9 in the CSF of dogs using gelatin zymography to verify possible alterations in these enzymes during natural systemic infection with Leishmania chagasi. Latent and active forms of MMP-2 were detected in some dogs of both groups, with high levels in the control group. In contrast, latent and active forms of MMP-9 were detected only in some animals with leishmaniasis. These results clearly demonstrate that MMP-9 is elevated in CSF of dogs with visceral leishmaniasis (VL). Although these results are preliminary, they suggest that MMP-9 might play a role in disruption of blood-brain barrier and/or blood-CSF barrier. While the presence of MMPs in CSF is not a condition exclusive to VL, their presence and persistence in CSF supports the hypothesis of an inflammatory state within CNS of dogs with VL.

Envelope or triangular flap for surgical removal of third molars? A systematic review and meta-analysis.

The aim of this systematic review was to compare the triangular and envelope flaps in mandibular third molar surgery with regard to pain, oedema, and trismus. Secondary outcomes assessed were dehiscence, ecchymosis, alveolar osteitis, periodontal condition, and surgical time. The PRISMA guidelines and recommendations in the Cochrane Handbook were followed, and the review was registered before commencement (PROSPERO; CRD42018112373). The literature search was conducted in the Web of Science, PubMed, Virtual Health Library, Cochrane Library, and Scopus databases and in the grey literature; randomized clinical trials, indexed through November 2018 were included. Three reviewers independently examined the studies. Twenty studies were included in the qualitative analysis, of which 18 were included in the meta-analyses. The flap design did not influence pain, oedema, trismus, dehiscence, or osteitis. The triangular flap was associated with a greater occurrence of postoperative ecchymosis (odds ratio 4.58, 95% confidence interval 1.34 to 15.91, I2=0) and lower periodontal probing depth on day 7 postoperative (standardized mean difference -1.36, 95% confidence interval -2.68 to -0.03, I2=88%) when compared to the envelope flap in mandibular third molar surgeries.

Leukocyte entry into the CNS of Leishmania chagasi naturally infected dogs.

In dogs, there is an association of chronic visceral leishmaniasis with neurological symptoms, and very few publications have investigated whether these neurological manifestations correlate with specific alterations in brain. A total of 42 mixed-breed adult dogs were selected from the Veterinary Hospital of UNESP-Araçatuba and the Control Zoonosis Center in Araçatuba, São Paulo State, Brazil, which is an endemic area for visceral leishmaniasis. Animals presenting positive ELISA and/or positive parasitological diagnosis of Leishmania were enrolled in the group of infected dogs (n=32). Animals with negative ELISA results and parasitological tests for Leishmania, including a negative immunofluorescence test for toxoplasmosis and neosporosis, were included as the control group (n=10). Brain samples were collected, stored in 10% buffered formalin and subjected to routine histological procedures, following by staining with haematoxylin-eosin (HE) and immunohistochemical examination for T and B lymphocytes and phagocytic cells. Cerebrospinal fluid was collected to determine the anti-Leishmania antibody titers. Histological examination of HE stains demonstrated intense inflammatory infiltrate, primarily in the choroid plexus, which was composed of mononuclear cells with no detectable parasites. Immunohistochemistry revealed that CD3(+) T lymphocytes were the major components of the inflammatory infiltrate at the choroid plexus and in the brain. Infected dogs had more CD3(+) T cells than uninfected animals (P=0.0002). Cerebrospinal fluid from infected dogs contained high titers of anti-Leishmania antibodies in comparison with control animals (P<0.0001), which suggests a compromise of the blood-cerebrospinal fluid barrier. Leukocyte entry into the brain suggests the participation of these cells in the pathogenesis of neurological disorders during the advanced stages of leishmaniasis and confirms that the choroid plexus is an important structure for T cell influx.

Epicarditis in a cat caused by feline infectious peritonitis virus: case report / Epicardite em um gato causada pelo vírus da peritonite infecciosa felina: relato de caso

Feline Infectious Peritonitis (FIP) is a progressive and fatal disease in domestic and wild cats, caused by Feline Infectious Peritonitis Virus (FIPV). The disease is characterized by an immunomediated reaction against the virus in various organs. This work described a case report of fibrinous epicarditis caused by FIPV. A male cat, three years old, died and was received to be necropsied. Grossly, soft, multifocal to coalescing, whitish fibrinous exudate, measuring up the 2 centimeters of thickness, was observed in the epicardium, mostly at the apex of the heart. Microscopically, severe, multifocal to coalescing inflammatory infiltrate was observed in the epicardium, composed mainly by macrophages, plasmocytes and lymphocytes, associated with fibrin deposition. Immunohistochemistry was performed for FIPV and was positive in the areas of inflammation in the epicardium. To the author´s knowledge, this is the second report of epicarditis due to FIPV in a cat. Therefore, epicarditis should be considered a differential diagnosis of cardiac diseases in Feline Medicine.(AU)

A Peritonite Infecciosa Felina (PIF) é uma doença progressiva e fatal de felinos domésticos e selvagens, causada pelo vírus da Peritonite Infecciosa Felina (FIPV). A doença é caracterizada por uma reação imunomediada contra o vírus em vários órgãos. Este trabalho descreveu um relato de caso de epicardite fibrinosa causada pelo FPIV. Um gato macho, com três anos de idade, veio a óbito e foi recebido para necropsia. Macroscopicamente, foi observado exsudato fibrinoso esbranquiçado, friável, multifocal, medindo até 2 centímetros de espessura, no epicárdio, principalmente no ápice cardíaco. Microscopicamente, foi observado no epicárdio um infiltrado inflamatório misto, multifocal, composto por macrófagos, plasmócitos e linfócitos, associado a deposição de fibrina. A imuno-histoquímica foi positiva para o FIPV nas áreas de inflamação no epicárdio. Os autores descrevem o segundo relato de caso na literatura científica de epicardite causada pelo FIPV em um gato. Portanto, a epicardite deve ser considerada como diagnóstico diferencial em doenças cardíacas em Medicina Felina.(AU)

Metaloproteinases no tecido laminar do casco de equinos submetidos à obstrução intraluminal do cólon menor / Metalloproteinases in hoof laminar tissue of equine submitted to intraluminal small colon obstruction

O objetivo deste estudo foi avaliar a expressão das MMP-2 e MMP-9 no tecido laminar do casco e o perfil leucocitário de equinos submetidos à obstrução intraluminal do cólon menor. Realizaram-se laparotomia e obstrução do cólon menor de oito equinos hígidos, utilizando-se uma bola inserida no lúmem intestinal. A bola foi inflada à pressão de 80mmHg e a obstrução foi mantida por quatro horas. Foram realizadas coletas sanguíneas antes da obstrução (M0), imediatamente após a desobstrução (M4) e a cada 12 horas após M4, até completar 72 horas (M12, M24, M36, M48, M60 e M72). As biópsias de casco foram realizadas em M0, M4 e M72, e as amostras foram submetidas à análise zimográfica. Foi observado aumento nos leucócitos em M12 e M24, decorrente do aumento de neutrófilos segmentados e bastonetes, os quais diminuíram a partir de M36. Segundo a técnica zimográfica, não se observaram alterações nos valores de MMP-2 e -9, possivelmente devido à baixa intensidade das lesões ocasionadas no cólon menor. Com isso, conclui-se que as alterações inflamatórias decorrentes da obstrução do cólon menor não foram suficientes para ocasionar alterações na expressão das MMP-2 e -9 no tecido laminar podal.(AU)

The aim of this study was to evaluate the blood leukocytes and the MMP-2 and -9 expression in the hoof laminar tissue of horses undergoing intraluminal small colon obstruction. Laparotomy and the small colon obstruction was performed in eight healthy horses, inserting a ball in the intestinal lumen. The ball was inflated to 80 mmHg pressure and the occlusion was maintained for 4 hours. The blood was collectedBlood samples were taken before the obstruction (M0), immediately after intestinal clearance (M4), and every 12 hours until completeuntil 72 hours (M12, M24, M36, M48, M60 and M72). The hoof biopsies were performed at M0, M4, and M72 and the samples were subjected to zymography analysis. There was an increase in leukocytes in M12 and M24, due to the increase in segmented neutrophils and band neutrophils, which decreased as of M36. According to zymography technique not observed changes were not not observed in MMP-2 and -9, possibly due to the low intensity of the small colon lesions. Wherefore, it is concludedIn conclusion, that the inflammatory changes resulting from small colon obstruction were not enough to cause changes in the expression of MMP-2 and -9 in the hoof laminar tissue.(AU)

Optic neuropathy in sheep associated with overdosage of closantel.

This report describes clinical and pathological findings in 2 flocks in Brazil where blindness and deaths in sheep occurred after closantel overdosage. Depression, weakness, and blindness affected 37 animals and 17 died in 2 flocks of 190 animals. Two animals submitted for ophthalmic examination showed no inflammation in the anterior segment of both eyes; posterior segment evaluation by indirect ophthalmoscopy suggested retinal degeneration. One postmortem evaluation local spongy vacuolization was in several regions of the brain and the optical nerves had severe axonal degeneration.

TNF-á and IL-6 immunohistochemistry in rat renal tissue experimentaly infected with Leptospira interrogans serovar Canicola

Leptospirosis is a public health problem worldwide and its etiology remains unclear. Its pathogenesis involves a complex interaction between host and infecting microorganism. The inflammatory reaction that controls the infection process also underscores many pathophysiological events occurring in leptospirosis. We investigated the presence of tumor necrosis factor-alfa (TNF-alfa) and interleukin-6 (IL-6) in renal tissues by immunohistochemical and histopathological examination in animals experimentally inoculated with Leptospira serovar Canicola. All the tests were carried out 2, 7, 14, 21 or 28 days after inoculation. Although TNF-alfa and IL-6 had been detected in tissues throughout the observation period, these cytokines appeared more intensely during the initial phase of infection. Therefore, both TNF-alfa and IL-6 were associated with the immunopathogenesis of leptospirosis. This profile suggests a high immunocellular response throughout the early infection stages followed by subsequent humoral response.

Vírus da estomatite vesicular (Sorotipo Indiana 2) como modelo experimental para o estudo de encefalite aguda: aspectos morfológicos / Vesicular stomatitis virus (Indiana 2 Serotype) as experimental model to study acute encephalitis: morphological features

O vírus da estomatite vesicular (VEV) é um vesiculovírus da família Rhabdoviridae que infecta mamíferos e causa lesões vesiculares semelhantes às observadas na febre aftosa. A encefalite experimental pode ser induzida em roedores e os sintomas são semelhantes aos observados na raiva; entretanto, as lesões observadas no encéfalo dos animais são diferentes. Corpúsculos de inclusão não são observados, há necrose especialmente da região do bulbo olfatório e em alguns casos, ventriculite. Observamos que o padrão temporal de disseminação do VEV e os aspectos morfológicos das lesões são similares aos descritos na literatura. O vírus parece se disseminar através dos ventrículos cerebrais, multiplicando-se em células do epêndima e em neurônios, além de utilizar o transporte retrógrado e anterógrado. Constatamos que devido à facilidade de manipulação do vírus, este modelo experimental tem sido utilizado em inúmeros trabalhos de pesquisa em diversas áreas. Se por um lado, os relatos sobre a patogenia da infecção são numerosos, por outro lado, ainda existem muitas lacunas que envolvem, por exemplo, aspectos sobre a transmissão do vírus, a recuperação dos animais infectados e a participação de células gliais durante a fase aguda e a fase de recuperação dos animais

Indicacoes cirurgicas de resseccao pulmonar na tuberculose da crianca. / Surgical indications for lung resection in tuberculosis of children

O autor estuda as indicacoes cirugicas atuais de resseccao pulmonar na tuberculose da crianca, apresentando dados referentes a idade, cor, evolucao, avaliacao pre-operatoria, complicacoes pos-operatorias imediatas, mortalidade e resultados, alem de focalizar detalhes de tecnica cirurgica, no que diz respeito a sutura do coto bronquico. Analisa sua casuistica de 43 casos operados, de criancas ate 12 anos, no periodo de janeiro de 1974 a janeiro de 1983, enfatizando as indicacoes atuais da resseccao pulmonar na doenca tuberculosa da crianca. Divide essas indicacoes em dois grupos (complicacoes da tuberculose primaria progressiva e doenca residual, apos tratamento da tuberculose de reinfeccao). Enfatiza a raridade de trabalhos sobre o assunto, na bibliografia nacional

Astrócitos imunorreativos à proteína glial fibrilar ácida (GFAP) em SNC de bovinos normais e de bovinos com raiva. I. Hipocampo e giro dentato / Glial fibrillary acidic protein (GFAP) immunoreactive astrocytes in the CNS of normal and rabies-infected adult cattle. I. Hippocampus and dentate gyrus

A proteína glial fibrilar ácida (GFAP) pertence ao grupo dos filamentos intermediários do citoesqueleto celular. Está presente nos astrócitos e em algumas células do epêndima. A identificaçäo da GFAP é empregada para identificar astrócitos com excelente especificidade. Neste trabalho, o método imunohistoquímico da peroxidase-antiperoxidase (PAP), utilizando anticorpo primário anti-GFAP, foi empregado para estudar a distribuiçäo e a morfologia de astrócitos no hipocampo e giro dentato de bovinos normais e também de bovinos com raiva, naturalmente adquirida. Foram utilizados cortes em parafina de material fixado em formalina tamponada. Astrócitos corados positivamente em marrom foram classificados em quatro níveis de intensidade. O número de astrócitos por mm2 foi determinado com uso de ocular integradora. Area, maior e menor diâmetro do núcleo dos astrócitos foram determinados com sistema analisador de imagens. Observou-se que bovinos com raiva mostraram astrogliose, apresentando forte imunorreatividade, aumento de tamanho do corpo celular e dos processos citoplasmáticos. Mostraram também astrocitose, sendo que, em todas as camadas do hipocampo e do giro dentato, o número de astrócitos foi maior que nos animais normais. O núcleo também se apresentava aumentado e pouco corado. Células nervosas da regiäo estudada näo apresentavam alteraçöes importantes. Os resultados colaboram para o conhecimento da distribuiçäo e da morfologia de astrócitos imunorreativos à GFAP, em bovinos normais e em bovinos com raiva